Fitness reduction for uncooperative fig wasps through reduced offspring size: a third component of host sanctions.

Mutually beneficial interactions between two species-mutualisms-are ancient, diverse, and of fundamental ecological importance. Nonetheless, factors that prevent one partner from reaping the benefits of the interaction without paying the cost are still poorly understood. Fig trees and their unique pollinators, fig wasps, present a powerful model system for studying mutualism stability. Both partners depend completely on each other for reproduction, cooperation levels can be manipulated, and the resulting field-based fitness quantified. Previous work has shown that fig trees can impose two types of host sanctions that reduce the fitness of wasps that do not pollinate: (1) fig abortion, which kills all developing larvae, and (2) reduced number of wasp offspring in figs that are not aborted. Here we demonstrate a third component of host sanctions. Through manipulative field experiments, we show that for four of five studied species, offspring of pollen-free foundresses are only 50-90% the size of offspring of pollinating foundresses. We further show that in all four studied species, smaller wasps are less likely to reach and enter a flowering fig to become foundresses themselves. Therefore, the experimentally determined size reduction of offspring is estimated to cause an additional reduction of up to 80% in fitness for a pollen-free foundress. We determine that the size reduction of pollen-free offspring acts on the level of the entire fig fruit rather than on individual flowers. These results show that estimates of the fitness effect of host sanctions on uncooperative symbionts should consider not only offspring quantity but also offspring quality. We discuss implications beyond the fig tree-fig wasp mutualism.

Population structure and the evolution of virulence in nematode parasites of fig wasps.

It is often assumed that parasitic and disease-producing organisms tend to evolve benign relationships with their hosts over time. In contrast, theoretical arguments suggest that increased opportunities for parasite transmission will promote the evolution of increased virulence. The natural history of species-specific nematodes that parasitize fig-pollinating wasps permits the testing of these predictions in natural populations. For 11 species of Panamanian fig wasps, those species characterized by population structures that result in increased opportunities for parasite transmission harbor more virulent species of nematodes. In addition, differences in population structure are also associated with differences in other intra- and interspecific phenomena, including sex ratios among the fig wasp species, the degree of tension in the wasp-fig mutualism, and lethal combat among the males of parasitic wasps.

Sex ratio adjustment in fig wasps.

The effects of inbreeding and local mate competition have often been confounded in theoretical and empirical studies of sex ratio. A general model considering the interplay of their influences on sex ratio evolution is presented, and its predictions are compared with data collected from three fig wasp species. The results indicate that female fig wasps adjust the sex ratio of their offspring in response to both the intensity of proximate local mate competition and the level of inbreeding in the population.

Evolution. The benefits of allocating sex.

Organisms allocate resources to male and female offspring in a process called sex allocation. In a Perspective, Stuart West and colleagues discuss what sex allocation tells us about evolution by natural selection and how sex allocation can be applied to understanding the mating structure of parasitic protozoans.

Genetic mosaics in strangler fig trees: implications for tropical conservation.

Single trees of six species of strangler figs (Ficus spp., Moraceae) in Panama were found to be made up of multiple genotypes, presumably formed by the fusion of different individuals. The phenomenon is frequent enough that strangler fig populations will contain considerably more genetic variation than would be expected from the number of trees. How this cryptic variation affects populations depends on the flowering phonology of composite trees. If the genetically different portions of trees flower asynchronously, populations of pollinating wasps may be more resistant to low host population sizes than previously thought. If different portions flower synchronously, attempts to infer mating-system parameters from the parentage of fruit crops will be misleading. The fruiting of figs, which are considered a keystone species in tropical forests, is important for maintaining biodiversity but is also particularly susceptible to failure at small population sizes. It is therefore important to know both the number of trees and the number of genotypes in a population.

Phylogenetic relationships, historical biogeography and character evolution of fig-pollinating wasps.

Nucleotide sequences from the cytochrome oxidase I (COI) gene were used to reconstruct phylogenetic relationships among 15 genera of fig-pollinating wasps. We present evidence supporting broad-level co-cladogenesis with respect to most but not all of the corresponding groups of figs. Using fossil evidence for calibrating a molecular clock for these data, we estimated the origin of the fig-wasp mutualism to have occurred ca. 90 million years ago. The estimated divergence times among the pollinator genera and their current geographical distributions corresponded well with several features of the break-up of the southern continents during the Late Cretaceous period. We then explored the evolutionary trajectories of two characteristics that hold profound consequences for both partners in the mutualism: the breeding system of the host (monoecious or dioecious) and pollination behaviour of the wasp (passive or active). The fig wasp mutualism exhibits extraordinarily long-term evolutionary stability despite clearly identifiable conflicts of interest between the interactors, which are reflected by the very distinct variations found on the basic mutualistic theme.

Physiological determinants of Ficus fruit temperature and implications for survival of pollinator wasp species: comparative physiology through an energy budget approach.

Figs are completely dependent for pollen dispersal on species-specific fig-pollinating wasps that develop within developing fig fruits. These wasps are very sensitive to heat and die at temperatures only a few degrees above ambient. Such temperatures are expected and observed in objects exposed to full sunlight, as fig fruits frequently are. In detailed field and experimental studies of 11 species of Panamanian figs with fruit ranging in size from 5 mm to 50 mm in diameter, we found that both the relative and absolute contribution of transpiration to maintaining non-lethal fruit temperatures increased with fruit size. Small and large fruits reached temperatures of 3 and 8°C, respectively, above air temperature in full sunlight when transpiration was prevented by grease. The temperature reached by large, nontranspiring fruits was sufficient to kill their pollinators. Control fruits which transpired reached temperatures of 2-3°C above air temperature in sunlight, regardless of size. An analysis of the solar energy budget of fruit revealed that large fruits must transpire to maintain tolerable temperatures for the wasps because heat diffusion from fruit to air was too low to balance net radiation in sunlight. By contrast, small fruits do not need to transpire to maintain tolerable temperatures for the pollinators.

Fruit characteristics and factors affecting fruit removal in a Panamanian community of strangler figs.

We describe fruiting characteristics for 12 species in a community of strangler figs (Moraceae: Urostigma) studied in Panama. We quantify diurnal and nocturnal removal rates and proportions of fruits removed, and relate them to the activities of the main dispersers of the figs: bats and birds. These results combined with previous studies show that there are clear differences between fig species with fruit that ripen red and those with fruit that remain green(ish). In the red-fruited species, the fruit are small, ripen asynchronously over relatively long periods, produce little scent, and are mainly taken during the day by birds. In contrast, in the green(ish)-fruited species, the fruits are larger, span a range of sizes, ripen relatively synchronously, produce very distinctive aromas, and are mainly taken at night by bats. This dichotomy in fruiting characteristics suggests coadaptive links between groups of dispersers and different species within the genus Ficus. All fig species produce a range of fruit crop sizes (10-155 fuits/m2 canopy area) of which a high proportion were removed by seed dispersers (>80%). Removal rates (fruit removed per day) were positively correlated with crop size, suggesting that trees with large crop size attract more frugivores. Removal rates of green-fruited figs were significantly lower and persistence and abortion of ripe fruit were significant higher around full moon, apparently due to the reduced activity of bats. We further estimate the number of bats that are sustained by a tree fruit crop and account for the observed fruit removal. We then discuss the evidence for coadaptation between different groups of figs and their seed dispersers, Finally, we consider the conservation implications for figs as keystone resources in tropical forests.

First Report of Phytophthora heveae and Pythium spp. on Tropical Tree Seedlings in Panama.

Damping-off was observed on experimentally planted seedlings of Ana-cardium excelsum (wild cashew), a timber tree, and Tetragastris panamensis, a canopy tree, within lowland tropical rain forest in Panama. Disease impact was greatest during the wet season (May through December). During the 1995 wet season, 40.7% (572/1,404) of T. panamensis seedlings died due to damping-off disease. Sixty-eight percent (703/1,034) of A. excelsum seed lings died due to damping-off during the 1996 wet season. Symptoms included leaf, cotyledon, and stem necrosis. Phytophthora heveae sporangia were observed on both host species, and oospores were found within stems of T. panamensis. Plating of diseased A. excelsum seedlings on potato dextrose agar with rifampicin (25 mg/ml) and pimaricin (10 mg/ml) produced cultures of Phytophthora heveae and Pythium from 27.4% (110/402) and 44.5% (179/402) of seedlings, respectively. Pythium isolates included P. vexans, P. splendens, and P. chamaehyphon species types, but P. vexans species types accounted for 70% of the Pythium isolates. Disease symptoms on experimental seedlings also were evident on naturally occurring seedlings. Mycelial plugs from six A. excelsum isolates of Phytophthora heveae were used to separately inoculate stems of three A. excelsum seedlings each. Of 18 seedlings inoculated, 88.8% developed characteristic symptoms and died in an average of 8.7 ± 1.0 (standard error [SE]) days. Nine Pythium isolates were used to separately inoculate stems of one to three A. excelsum seedlings each; three of these isolates were known to be P. vexans species types. All of the 20 seedlings inoculated with a Pythium isolate developed characteristic symptoms and died in an average of 6.1 ± 0.3 (SE) days. Both Phytophthora heveae and Pythium isolates were reisolated readily from diseased seedlings. Cotyledons and stems of seven to eight T. panamensis seedlings per isolate were inoculated with two Phytophthora heveae isolates originating from T. panamensis. Necrotic lesions on cotyledons consistent with field symptoms developed on 33.3% of 15 seedlings, but disease did not spread within the stem. Measurements of key morphological structures and cardinal temperatures of four Phytophthora heveae isolates from A. excelsum were consistent with published species descriptions (1), except (i) sporangia with two apices were present, although infrequent; (ii) chlamydospores were produced; and (iii) antheridia were narrower and often shorter than published measurements (7 to 12 m long; 2 to 6 m wide). Internal transcribed spacer (ITS) sequences from Phytophthora heveae isolates cultured from A. excelsum and T. panamensis were matched to reference sequences of Phytophthora heveae with only 3-bp differences (2). ITS sequences for isolates of Pythium vexans, P. splendens, and P. chamaehyphon species types clustered within clades of reference strains of these species (C. A. Lévesque, personal communication). Phytophthora heveae and Pythium spp. have been reported from the tropics. However, this is the first report of these pathogens on seedlings of A. excelsum and T. panamensis. Reference: (1) D. C. Erwin and O. K. Ribeiro. 1996. Phytophthora Diseases Worldwide. The American Phytopathological Society, St. Paul, MN, pp. 100-107, 336-337. (2) D. E. L. Cook et al. A molecular phylogeny of Phytophthora and related Oomy-cetes. Fungal Genet. & Biol. In press.

Histological Comparisons of Parasitism by Schistonchus spp. (Nemata: Aphelenchoididae) in Neotropical Ficus spp.

Syconia (enclosed infructescences) infested with host-specific species of Schistonchus (Aphelenchoididae) were collected from six species of Ficus (Moraceae) native to Florida or Panama. They were sectioned and histologically examined to assess the effects of parasitism. Parasitism by Schistonchus spp. was associated with hypertrophied cells, tissue necrosis, and the presence of an exudate in all species. Occasional hypertrophy of the outer epidermal cells occurred on seed florets, wasp florets, and on the endothecial cells of male florets in F. aurea (subgenus Urostigma) from Florida. Aberrations of the inner mesocarp occurred under the hypertrophied cells on seed florets. In F. laevigata (subgenus Urostigma) from Florida, Schistonchus sp. infested immature male florets and was associated with hypertrophy of endothecial cells, epidermal cells of the anther filaments, and anthers. Schistonchus sp. also caused aberrations of the anther filament, anthers, and pollen. Ficus poponoei (subgenus Urostigma) and F. glabrata (subgenus Pharmacosycea), both from Panama, had hypertrophied outer epidermal cells on seed florets. Ficus poponoei also had Schistonchus sp. within the pedicel of an aborted floret, with hypertrophy of the cortical parenchyma. Ficus trigonata (subgenus Urostigma) from Panama had hypertrophy of the outer epidermis of seed florets. When the outer epidermis on these florets was missing, the inner mesocarp was hypertrophied. Ficus maxima (subgenus Pharmacosycea) from Panama had hypertrophy on the outer epidermis of seed and aborted florets. Schistonchus spp. were not found in wasp larvae or pupae in any of the Ficus spp. examined. Hypertrophy was never observed in the absence of Schistonchus spp.

The evolution of parasitic diseases.

Parasites are characterized by their fitness-reducing effect on their hosts. Studying the evolution of parasitic diseases is an attempt to understand these negative effects as an adaptation of the parasite, the host, both or neither. Dieter Ebert and E. Allen Herre here discuss how the underlying concepts are general and are applicable for all types of disease-producing organisms, broadly defined here as parasites. The evolutionary processes that lead to the maintenance of the harmful effects are believed to be characterized by genetic correlations with other fitness components of the parasite. Depending on the shape of these correlations, any level of virulence can evolve.

STABILIZING SELECTION AND VARIANCE IN FIG WASP SEX RATIOS.

Theory predicts that the phenotypic variance observed in a trait subject to stabilizing selection should be negatively correlated with the trait's impact on fitness. However, this relationship has rarely been tested directly. The offspring sex ratios produced by pollinating fig wasp foundresses upon entrance to a fruit and oviposition alone (single foundress sex ratios) are subject to stabilizing selection because too many males reduce the total number of dispersing females and too few males will result in unmated females or complete loss of the brood. Furthermore, we argue that the impact on fitness of, and therefore the intensity of stabilizing intensity on, single foundress sex ratios are correlated to how frequently a species produces single foundress broods in nature. Specifically, the intensity of stabilizing selection will be greater in species that encounter single foundress broods more frequently, both because the trait is expressed more often and because fitness shows a greater sensitivity to variation (narrower fitness profile) when that trait is expressed. Across 16 species of Panamanian pollinating fig wasps, the phenotypic variance in single foundress sex ratios was negatively correlated with the frequency with which that species encounters single foundress broods in nature. In addition, a formal comparative analysis based upon a molecular phylogeny of the wasps gave results that were the same as when species were used as independent data points.

Pervasive density-dependent recruitment enhances seedling diversity in a tropical forest.

Negative density-dependent recruitment of seedlings, that is, seeds of a given species are less likely to become established seedlings if the density of that species is high, has been proposed to be an important mechanism contributing to the extraordinary diversity of tropical tree communities because it can potentially prevent any particular species from usurping all available space, either in close proximity to seed sources or at relatively larger spatial scales. However, density-dependent recruitment does not necessarily enhance community diversity. Furthermore, although density-dependent effects have been found at some life stages in some species, no study has shown that density-dependent recruitment affects community diversity. Here we report the results of observations in a lowland, moist forest in the Republic of Panamá in which the species identities of 386,027 seeds that arrived at 200 seed traps were compared with the species identities of 13,068 seedlings that recruited into adjacent plots over a 4-year period. Across the 200 sites, recruit seedling diversity was significantly higher than seed diversity. Part of this difference was explained by interspecies differences in average recruitment success. Even after accounting for these differences, however, negative density-dependent recruitment contributes significantly to the increase in diversity from seeds to seedling recruits.

Molecular diversity of arbuscular mycorrhizal fungi and patterns of host association over time and space in a tropical forest.

We have used molecular techniques to investigate the diversity and distribution of the arbuscular mycorrhizal (AM) fungi colonizing tree seedling roots in the tropical forest on Barro Colorado Island (BCI), Republic of Panama. In the first year, we sampled newly emergent seedlings of the understory treelet Faramea occidentalis and the canopy emergent Tetragastris panamensis, from mixed seedling carpets at each of two sites. The following year we sampled surviving seedlings from these cohorts. The roots of 48 plants were analysed using AM fungal-specific primers to amplify and clone partial small subunit (SSU) ribosomal RNA gene sequences. Over 1300 clones were screened for random fragment length polymorphism (RFLP) variation and 7% of these were sequenced. Compared with AM fungal communities sampled from temperate habitats using the same method, the overall diversity was high, with a total of 30 AM fungal types identified. Seventeen of these types have not been recorded previously, with the remainder being similar to types reported from temperate habitats. The tropical mycorrhizal population showed significant spatial heterogeneity and nonrandom associations with the different hosts. Moreover there was a strong shift in the mycorrhizal communities over time. AM fungal types that were dominant in the newly germinated seedlings were almost entirely replaced by previously rare types in the surviving seedlings the following year. The high diversity and huge variation detected across time points, sites and hosts, implies that the AM fungal types are ecologically distinct and thus may have the potential to influence recruitment and host composition in tropical forests.

Testing Hamilton's rule with competition between relatives.

Hamilton's theory of kin selection suggests that individuals should show less aggression, and more altruism, towards closer kin. Recent theoretical work has, however, suggested that competition between relatives can counteract kin selection for altruism. Unfortunately, factors that tend to increase the average relatedness of interacting individuals--such as limited dispersal--also tend to increase the amount of competition between relatives. Therefore, in most natural systems, the conflicting influences of increased competition and increased relatedness are confounded, limiting attempts to test theory. Fig wasp taxa exhibit varying levels of aggression among non-dispersing males that show a range of average relatedness levels. Thus, across species, the effects of relatedness and competition between relatives can be separated. Here we report that--contrary to Hamilton's original prediction but in agreement with recent theory--the level of fighting between males shows no correlation with the estimated relatedness of interacting males, but is negatively correlated with future mating opportunities.